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Potentiometric determination of α-l-fucosidase enzyme by using 2-chloro-4-nitrophenol-rhodamine B ion pair chemical recognition in PVC membrane sensor

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Abstract

The activity of the α-l-fucosidase (AFU) enzyme represents an excellent test for diagnosis of hepatocellular carcinoma (HCC) and fucosidosis recognized in inborn disorder of metabolism and increases the sensitivity of detection to 95.5% in patients with HCC. Therefore, the determination of the activity of AFU enzyme is very important and can be used as a screening tool for the early diagnosis of tumors for HCC patients. A simple, accurate, and sensitive potentiometric method was developed for measuring the activity of AFU. The method was based upon measuring the concentration of 2-chloro-4-nitrophenol (2-chloro-4-NP) using a 2-chloro-4-NP-rhodamine B ion pair in a PVC membrane sensor. The electrode shows a linear, reproducible, and stable potentiometric response with an anionic Nernstian slope of −51.13 ± 0.6 mV/decade over a wide range of concentrations 10−5–10−2 M and a detection limit of 1.0 × 10−6 M of 2-chloro-4-NP. The membrane exhibits a fast response time of 30 s, over a pH range of 4.0–6.5. The selectivity coefficients indicate excellent selectivity for 2-chloro-4-NP over a number of interfering species, e.g., chloride, nitrate, sulfate, chromate urea, albumin, glucose, uric acid, and total protein. The prepared sensor has been used successfully for the determination of 2-chloro-4-NP produced from the hydrolysis of 2-chloro-4-NP-α-l-fucopyranoside substrate. It was also applied for the determination α-l-fucosidase enzyme of 33 serum samples of healthy subjects and patients. The average recoveries ± RSD for the healthy subjects, cirrhosis of chronic hepatitis C and B, and HCC serum samples were 102.6 ± 1.01%, 101.5 ± 0.95%, and 100.1 ± 1.1%, respectively. The results obtained are in good agreement with those obtained by standard methods.

Potentiometric response of 2-chloro-4-NP using 2-chloro-4-NP-Rd PVC membrane sensor in phosphate buffer of pH 5 at 25 °C

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Authors and Affiliations

  1. Genetic Engineering and Biotechnology institute, Menoufia University, Sadat City, 32551, Egypt

    Abdelhameed M. Othman

  2. Nation cancer Institute, Cairo University, Cairo, 11562, Egypt

    M. E. El-Houseini

  3. Faculty of Engineering, Modern Academy University, Cairo, 11357, Egypt

    Mohamed S. El-Sofy

  4. Pharmaceutical and Medicinal Chemistry Department, National Research Centre, Dokki, Cairo, 12311, Egypt

    Hassan Y. Aboul-Enein

Authors
  1. Abdelhameed M. Othman
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  2. M. E. El-Houseini
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  3. Mohamed S. El-Sofy
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  4. Hassan Y. Aboul-Enein
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Correspondence to Abdelhameed M. Othman or Hassan Y. Aboul-Enein.

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Othman, A.M., El-Houseini, M.E., El-Sofy, M.S. et al. Potentiometric determination of α-l-fucosidase enzyme by using 2-chloro-4-nitrophenol-rhodamine B ion pair chemical recognition in PVC membrane sensor. Anal Bioanal Chem 400, 787–795 (2011). https://doi.org/10.1007/s00216-011-4774-0

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  • DOI: https://doi.org/10.1007/s00216-011-4774-0

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